PROJECT SUMMARY (Overall Unit) Metastasis embodies the dynamic, complex, whole-organism pathophysiology of cancer. Spread of cancer cells beyond the primary tumor site is the cause of the majority of cancer deaths and the most overt expression of cancer's complex evolutionary dynamics. Intimately related to the intricate processes of development and immunity, the transition from locally invasive to metastatic cancer poses a major scientific hurdle. Recent technological and computational advancements, pioneered by our team enable dynamic, multi-dimensional, multi-planar analysis of multiple tissues. Applying these advances to the most lethal cancers in the US: Lung Cancer, Pancreatic Cancer and CNS metastases, we will generate a Human Tissue Atlas with unprecedented spatial-temporal resolution. This multidisciplinary effort begins with an unparalleled array of human tissues and clinical data acquired by our Biospecimen Unit team. These high-quality biospecimens will be obtained from lung cancer, pancreatic cancer and CNS metastases obtained from surgical resections, biopsy or autopsy. The Data Analysis Unit will then subject these specimens will be subjected to multi-dimensional analysis including bulk DNA sequencing, single cell and single nuclear RNA sequencing, as well as architecturally- sensitive analyses including MIBI (protein) and MERFISH (RNA) technologies. Driven by revolutionary computational and systems biologists, integration and presentation of these datasets will be undertaken by the Characterization Unit, with the goal of generating authoritative human tumor atlases of value to the entire cancer research community.